HU198723B - Process for producing macrolide compounds and pharmaceutical compositions comprising such compounds as active ingredient, and plant protectives comprising such compounds - Google Patents

Description

The present invention relates to novel macrolide compounds and to pharmaceutical compositions containing such compounds. Compositions containing the compounds of the present invention have valuable acaricidal, insecticidal, and antifungal activity, which are generally referred to as anti-parasitic activities. The invention also relates to pesticides containing such compounds.

The novel compounds of the present invention are chemically related to known milbemycins and avermectins, among others. Known compounds having a 16-membered macrolide ring structure based There are several classes ^IS-13-size, which are obtained semi-synthetically produced by fermentation or by chemical derivatization of such natural fermentation products of various microorganisms and which exhibit acaricidal, anthelmintic and other antiparasitic activities. Milbemycins and avermectins can be considered as two distinct groups of such known compounds, but there are many other known compounds identified by different names or code numbers. The names of these various macrolide compounds are generally taken from the name or code number of the microorganisms capable of producing naturally occurring members of these groups, and these names are usually used to include all chemically related compounds of the same class. . As a result, there is no standardized systematic nomenclature that would apply to these compounds in general.

To avoid any misunderstanding, the following will use a standardized nomenclature system based on the hypothetical parent compound represented by the formula ™ (A). For the avoidance of doubt, Formula (A) is numbered at the positions of the macrolide ring system which are most relevant to the compounds of the invention described below. The carbon atom of the 4-position methyl group is given the number 26.

Thus, naturally occurring milbemycins are a group of macrolide compounds. A ₃ and A 4 in ⁵⁰ milbemycins 3,950,360. U.S. Pat. A milbemycin D a

4346171. U.S. Pat. These compounds may also be represented by the above formula (A), wherein the 25-position of these compounds is substituted by methyl, ethyl or isopropyl, respectively, as milbemycin A ₃ , milbemycin A4 or milbemycin D. The milbemycin analog substituted at the 25-position with a sec-butyl group is disclosed in U.S. Pat. U.S. Pat.

Subsequently, various derivatives of the original milbemycins were prepared and the effect of these derivatives was investigated. Milbemycins esterified at the 5-position are described in

4.2018, 861, 4,206,205, 4,173,571, 4,111,314,

4,293,976., 4,289,760., 4,457,920., 4,579,864. and

4547491. U.S. Patent Nos. 8,184, 102,721, U.S. Pat.

115,930., 142,969., 180,539. and 184,989. European Patent Publication Nos. 57-120589 and 59-168894.

No. 4,423,209. 13-hydroxy-5-oxo-milbemycin derivatives have been disclosed in US patent no. A 4.547.520. and U.S. Patent No. 203,832. European Patent Publication No. 4,123,125 discloses milbemycin-5-oxime derivatives. A 2.168.345. United Kingdom Patent No. 5,123,015 discloses milbemycin derivatives having a carboxyl or esterified carboxyl group at the 13-position in combination with a hydroxyl or esterified hydroxyl group.

Like milbemycins, avermectins can be derived from the same macrolide compound containing a 16-membered ring. Avermectins are described, for example, in J. Antimicrob. Agents Chemother., 15 (3), 361-367 (1979) and J. Am. Chem. Soc., 103, 4216 (1981). These compounds are represented by the above formula (A) but with the 4 '- (alpha-L-oleandrosyl) -alpha-L-oleandrosyloxy group at the 13-position. The 25-position may be replaced by an isopropyl group or a sec-butyl group, and / or a carbon-carbon double bond is present between the 22- and 23-positions or a hydroxyl group is attached at the 23-position. Avermectins can be defined as follows:

avermectin ^C 22 ~ ^C 23 ^R 25 ^R 23 ^r 5 A | 3 kk sec-Bu H OMe A] b kk i-Pr H OMe _2a wedge sec-Bu OH OMe In _{Fig. 2} b wedge i-Pr OH OMe - B _t a kk sec-Bu H OH B] b kk i-Pr H OH B ₂ a wedge sec-Bu OH OH B ₂ b wedge i-Pr OH OH

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The above Table 25 substituent, R ₂ 3 23 5-substituent and R5 substituent, mk indicates the double bond between 22- and 23-positions and s represents a single bond between the 22- and 23-positions.

Avermectin A ₂ a, A ₂ b, B ₂ a and B ₂ b 23-oxo-derivates are disclosed in U.S. Pat. U.S. Pat. 22,23-Dihydro-avermectins may be prepared by reduction of the double bond between the 22 and 23 positions and the 4,199,569. U.S. Pat. Aglucone derivatives of avermectins (which are milbemycin analogs) are sometimes referred to in the literature as C-076 compounds, and various derivatives of these compounds are known. For example, U.S. Pat. U.S. Pat. No. 5,198 discloses derivatives substituted at the 13-position by a lower alkanoyl group.

A 74,758. European Patent Application EP-A-004123 discloses avermectin derivatives which are formed on the 4-position methyl group. Thus, in this specification, the conversion of 4-methyl to hydroxymethyl is described, together with the preparation of a wide variety of oxymethyl derivatives such as acetyloxymethyl, benzoyloxymethyl and other carbonyloxymethyl derivatives.

A 170.006. European Patent Application Publication No. 5,123,121 discloses a family of bioactive compounds produced by fermentation, collectively identified by the code number LL - F28249. Some of these compounds have a 16-membered macrolide having the structure (A) above, with hydroxy at position 23 and 1-methyl-1-propenyl, 1-methyl-1-butenyl or 1 , Substituted with 3-dimethyl-1-butenyl. In these compounds, the 5-hydroxy group may be replaced by a methoxy group.

No. 2,166,436. U. S. Patent No. 5,481,121 discloses compounds of the same or similar designations as S-541. The 23-oxo and 23-deoxy derivatives of S-541 are disclosed in U.S. Patent No. 904,709. Belgian Patent No. 5,198,198. European Patent No. 215,654 discloses S-541 derivatives having a carbon-carbon double bond between the 22 and 23 positions. A 237.341. European Patent Application No. S-541 discloses 26-hydroxy and 26 (C 1 -C 4) -alkanoyloxy derivatives, as well as 23-oxo and 23-deoxy derivatives.

In 2.176.182. Another published group of macrolide antibiotics corresponding to the above formula (A) is known from the published British Patent Application Publication No. 5,103, optionally substituted hydroxy or oxo at position 5, and alpha-branched alkenyl at position 25. Including.

A further group of related macrolide derivatives is disclosed in Japanese Patent Application Laid-Open No. 62-29,590. The structure of these compounds corresponds to the structure (A) above, having a hydroxy or methoxy group at the 5-position. It is substituted at the 13-position of the ring with a 4 '- (alpha-Loleandrosyl) -alpha-L-oleandrosyl] oxy group, as in the case of avermectins, and may be carbon-carbon double bonded between the 22 and 23 positions or alternatively the 23-position may be replaced by a hydroxyl group. The 25-position substituent is of a type not found in naturally occurring avermectins and micromycins. For example, this substituent may be various alpha-branched alkyl, alkenyl, alkynyl, alkoxyalkyl, alkylthioalkyl and cycloalkylalkyl groups, or cycloalkyl, cycloalkenyl or heterocyclic groups. This 25-substituent is introduced into the molecule by adding the appropriate carboxylic acid or a reactive derivative thereof to the fermentation broth obtained by culturing the avermectin-producing microorganism.

All of the above-described milbemycin-related macrolide compounds are known to possess one or more of the antibiotic, anthelmintic, ectoparasiticidal, acaricidal, or other pesticidal effects. However, there is a continuing need for compounds that exhibit modified activity in one or more classes of pests.

It has been found that the activity of the milbemycin derivatives can be modified by introducing an unsaturated ester group at the 4-position to give the 26-alkenyloxy derivatives of the milbemycins and related macrolides.

The present invention therefore relates to a process for the preparation of macrolide compounds of formula (I)

-XY- represents -XY- represents -CH ₂ -CH ₂ - or CH ₂ -CH ₂ H-,

R ¹ is methyl, ethyl, isopropyl, sec-butyl or 'C (CH 3) = CHR ⁵ and in the latter R ³ is methyl, ethyl or isopropyl,

R ² is - (CH 2) n C) R ⁶ ) = C (R ⁷ ) (R ⁸ ) wherein n is 0, 1 or 2, R ⁶ and R ⁷ independently of one another are hydrogen or methyl; R ⁸ is hydrogen or C 1-4 alkyl, or phenyl optionally substituted with halo, provided that when R ⁸ is C 1-4 alkyl, then - (CH 2) _n -C (R ⁶ ) = C ( R ⁷ ) The alkenyl group of the formula (R ⁸ ) contains up to 8 carbon atoms,

R ³ is hydrogen, C 2 -C 5 alkanoyl, C 2 -C 5 alkoxycarbonyl, or tri (C 1 -C 4 alkylsilyl) and

R ⁴ is hydrogen or alpha-L-oleandrosyl-alpha-L-oleandrosyloxy, with the proviso that R ⁴ is hydrogen when R ¹ is methyl, ethyl or -C (CH ₃ ) "CHR ³

198723 Β and in the latter R ^J is methyl, ethyl or isopropyl.

The invention further relates to a process for the production of anti-parasitic compositions having, for example, an antifungal, acaricidal or insecticidal activity. The formulations may contain, in addition to the active ingredient of formula (I), a carrier or diluent for pharmaceutical, agricultural, veterinary or horticultural use and other excipients.

The compositions of the invention may thus be used to treat or protect humans, animals and plants to eliminate or prevent damage by parasites such as mites, worms and insects, for example by administering the composition to or in the vicinity of an infected human, animal, plant or plant seed. .

Returning to Formula I, when R ² is -C (CH 3) = CHR ⁵ , R ^{2 is} more particularly 1-methyl-1-propenyl,

It may be 1-methyl-1-butenyl or 1,3-dimethyl-1-butenyl.

When R ³ is a trialkylsilyl group, examples of such silyl groups include trimethylsilyl, triethylsilyl, tripropylsilyl, triisopropylsilyl, diisopropylmethylsilyl and tert-butyldimethylsilyl. . Of these groups, trimethylsilyl, triethylsilyl and tert-butyldimethylsilyl are preferred.

Preferred are compounds of formula I wherein (a) -XY- is -CH ₂ -CH ₂ - or -CH ₂ -CHOH, (b) (i) R 'is methyl, ethyl or isopropyl and R ⁴ is hydrogen, or (b) (ii) ^R1 is isopropyl or sec-butil10 group and R ⁴ is alpha-L-oleandrosyl-alpha-L-oleandrosyl group, or (b) (iii) R ¹ is -C (CH ₃ ) = CHR ⁵ (wherein R ⁵ is as previously defined) and R ^{4 is} hydrogen, (c) R ² is -CH = C (CH ₃ ) (R ^10). ) wherein R ^{10 is} methyl or ethyl, (d) R ³ is hydrogen, alkanoyl having 2 to 4 carbon atoms (such as acetyl or propionyl) or alkoxycarbonyl having 2 to 5 carbon atoms (e.g. ethoxy). -carbonyl).

Particularly preferred compounds are those compounds of formula (I) wherein -XY- is CH ₂ -CH ₂ -, R ¹ is methyl or elyl, R ² is 2-methyl-1-propenyl, and R ³ and R ⁴ are hydrogen. .

Among the compounds of formula (I), preferred compounds are listed in the following table:

The compound number X-Y ^R 1 r ₂ r ₃ r ₄ First -ch ₂ -ch ₂ Me -CH = C (CH ₃ ) ₂ H H Second -ch ₂ -ch ₂ Me - CH = C (CH ₃ ) (C ₂ H ₅ ) H H Third -ch ₂ -ch ₂ et -CH = C (CH ₃ ) ₂ H H 4th -ch ₂ ch ₂ et - CH = C (CH ₃ ) (C ₂ H ₅ ) H H 5th -ch ₂ -ch ₂ et -ch = chch ₃ H H 6th -ch ₂ -ch ₂ et -ch = ch ₃ ) = chch ₃ H H 7th -ch ₂ -ch ₂ et -CH == CH (CH ₂ ) ₂ CH ₃ H H 8th -ch ₂ -ch ₂ et -CH = CHC (CH ₃ ) ₂ H H 9th -ch ₂ -ch ₂ et -ch ₂ ch = chc ₂ h ₅ H H 10th -ch ₂ -ch ₂ et -ch ₂ ch ₂ ch = ch ₂ H H 11th -ch ₂ -ch ₂ et - CH = CH-Phenyl H H 12th -ch ₂ -ch ₂ et - CH * = CH-4-chlorophenyl H H 13th -ch ₂ -ch ₂ et —CH = C (CH ₃ ) ₂ BMS H 14th -ch ₂ -ch ₂ et - CH = C (CH ₃ ) ₂ COC ₂ H ₅ H 15th -ch ₂ ch ₂ et -CH = C (CH ₃ ) ₂ COOC ₂ H ₃ H 16th -ch ₂ -ch ₂ iPr -CH = C (CH ₃ ) ₂ H H 17th -ch ₂ -ch ₂ sec-Bu -CH ₂ C (CH ₃ ) ₂ H alpha 18th -CH = CH- sec-Bu -CH = C (CH ₃ ) ₂ H alpha 19th -CH ₂ -CH ₂ DMB -CH-C (CH ₃ ) ₂ H H 20th -ch ₂ -choh- DMB -CH-C (CH ₃ ) ₂ H H 21st -CH ₂ -C (»O) - DMB -CH ₂ C (CH ₃ ) ₂ H H

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In this table:

BMS: tert-butyldimethylsilyl, alpha: alpha-L-oleandrosyl-alpha-L-oleandrosyloxy,

DMB; 1,3-dimethyl-l-butenyl.

The compounds listed in the table are named as follows:

26- (3-Methyl-2-butenoyloxy) -milbemycin A ₃

2. 26- (3-Methyl-2-pentenoyloxy) -milbemycin ^A 3

3. 26- (3-Methyl-2-butenoyloxy) -milbemycin A ₃

4. 26- (3-Methyl-2-pentenoyloxy) -milbemycin ^A 3

5. 26- (2-Butenoyloxy) -milbemycin A4

6. 26- (2-Methyl-2-butenoyloxy) -milbemycin A4

7. 26- (2-Hexenoyloxy) -milbemycin A4

8. 26- (4-Methyl-2-pentenoyloxy) -milbemycin A4

9. 26- (3-Hexenoyloxy) -milbemycin A4

10. 26- (4-Pentenoyloxy) -milbemycin A4

11. 26- (Cinnamoyloxy) -milbemycin A4

12. 26- (4-Chloro-cinnamoyloxy) -milbemycin A4 • 13, 5- (O-tert-Butyl-dimethylsilyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4

14. 5- (O-Propionyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4

15. 5- (O-Ethoxycarbonyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4

16. 26- (3-Methyl-2-butenoyloxy) -milbemycin D

17. 26- (3-Methyl-2-butenoyloxy) -ivermectin B1a

18. 26- (3-Methyl-2-butenoyloxy) avermectin B1a

19. 23-Deoxy-26- (3-methyl-2-butenyloxy) -S-541A

20. 26- (3-Methyl-2-butenoyloxy) -S-541A

21. 23-Oxo-26- (3-methyl-2-butenoyloxy) -S-541A.

Of these compounds, compounds 1, 2, 3, 4, 7, 8, 10, 11, 14, 15, 16, 17, 18, 19, 20 and 21 are preferred and particularly preferred are 1, 2, 3, 4, 16, 17, 18, 19, 20 and 21.

The compounds of formula (I) can be prepared in steps A and B as shown in Scheme 1. In Scheme 1, -XY and R ¹ -R ⁴ are as previously defined.

Thus, in step A, the 4-methyl group, i.e. the methyl group attached at the 4-position of the milbemycin molecule, is subjected to selective oxidation and is thus converted to the 4-hydroxymethyl group. This reaction is known as Sharpless reaction or Sharpless oxidation and can be understood as allyl oxidation with selenium dioxide and tert-butyl hydroperoxide. The Sharpless reaction has been used previously for macrolides of formula II as a type. In this regard, reference is made to European Patent No. 74758 and U.S. Patent No. 237,341. European Patent Publication No. 3,600,198. Step A can thus be performed under conditions similar to those described in these specifications.

Thus, in Step A, the 4-methyl group of a compound of formula II is oxidized with tert-butyl hydroperoxide in the presence of catalytic amount of selenium dioxide which oxidizes the 4-methyl group to a 4-hydroxymethyl group. The tert-butyl hydroperoxide re-oxidizes the reduced selenium compound to selenium dioxide for further oxidation. In this way, only a small, catalytic amount of selenium dioxide is required.

The reaction is carried out in an inert solvent which is not sensitive to oxidation. Methylene chloride is preferred, although ethyl acetate, tetrahydrofuran and other solvents may also be used. The reaction temperature is not critical, for example, from 0 ° C to 50 ° C. Preferably, room temperature is employed. The reaction is generally complete within 1 to 48 hours and, under the preferred reaction conditions, typically about 24 hours is sufficient

For compounds of formula II, R ^{3 is} preferably a methyl, hydroxy protecting group, or ester-forming carboxylic acid or carbonic acid moiety that functions as a protecting group. When R ^{3 in} the compound of formula (II) and thus in the intermediate of formula (III) is hydrogen, it is preferred to convert this hydrogen atom to one of the said groups before carrying out step B by one of the methods described above.

In step B of the 26-hydroxyl group of the intermediate of formula (III) with a carboxylic acid of formula R ² COOH - wherein ^R2 is as defined above - or a reactive derivative thereof, in a known manner.

Reactive derivatives of carboxylic acids of the formula R ² COOH include acid halides such as acid chloride, acid bromide or acid iodide; acid anhydride; mixed acid anhydrides; reactive esters such as pentachlorophenyl esters or

4-nitrophenyl esters; and reactive acid amides.

Step B is preferably carried out in the presence of a solvent. The solvent quality is not critical provided that the reaction is not adversely affected. Suitable solvents include aliphatic or aromatic, optionally halogenated hydrocarbons such as hexane, petroleum ether, benzene, toluene, xylene, chloroform, methylene chloride or chlorobenzene; ethers such as diethyl ether, tetrahydrofuran or dioxane; and esters such as methyl acetate or ethyl acetate. The reaction is carried out over a wide range of temperatures, and the particular temperature employed is not critical. The reaction time may vary depending on a number of factors, but generally between 30 minutes and 3 hours may be sufficient.

Generally, 1 to 10 molar equivalents of acid or reactive derivative thereof are used per mole of compound of formula III, more preferably 1.5 to 4 molar equivalents.

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When the acid itself is used, the reaction is preferably carried out in the presence of a dehydrating agent such as dicyclohexylcarbodiimide (DCC), ethyl polyphosphate (PPE), mesitylene sulfonyl triazolid (MST), ptoluenesulfonic acid or sulfuric acid, particularly DCC. The amount of dehydrating agent is usually 1 to 5 molar equivalents, preferably 1.5 to 4 molar equivalents. When DCC is used, the reaction is conveniently carried out in a catalytic amount of a base such as pyridine or

Is carried out in the presence of 4-pyrrolidinopyridine.

When a reactive derivative of the acid is used, the reaction is preferably a base, particularly an organic base such as triethylamine, Ν, Ν-dimethylaniline, pyridine, 4- (dimethylamino) pyridine, 1,5-diazabicyclo 4,3.0/non-5-ene (DBN) or 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU). The base is usually used in an amount of 2 to 8 molar equivalents.

When the reaction product of Step B carries a silyl group as a protecting group, it can be readily removed to convert it to the 5-hydroxy group. The removal may be carried out with a dilute mineral acid such as dilute hydrochloric acid or dilute sulfuric acid; an organic acid such as formic acid or acetic acid; using a mixture of p-toluenesulfonic acid and letrabutylammonium fluoride or a mixture of hydrogen fluoride and pyridine. Of these, dilute hydrochloric acid and a mixture of hydrogen fluoride and pyridine are preferred. The acid is usually used in an excess, preferably in an amount of 2 to 100 molar equivalents.

The deprotection is preferably carried out in the presence of a solvent. The solvent quality is not critical provided that the reaction is not adversely affected. Suitable solvents include aliphatic or aromatic and optionally halogenated hydrocarbons such as hexane, petroleum ether, benzene, toluene, chloroform, methylene chloride or dichloroethane; ethers such as dictylether, tetrahydrofuran, dioxane or dimethoxyethane; alcohols such as methanol or ethanol; nitriles such as acetonitrile or propionitrile; and mixtures of these solvents.

The reaction takes place over a wide range of temperatures and thus the exact temperature is not critical. However, in general, we find it convenient to carry out the reaction at a temperature of -20 ° C to + 70 ° C, more preferably at a temperature of -10 ° C to + 30 ° C. The reaction time may vary depending on many factors; however, a period of from 30 minutes to 24 hours, preferably from 30 minutes to 6 hours, is usually sufficient.

The compounds of formula I wherein R ³ is hydrogen may, if desired, be subsequently esterified with a carboxylic acid or carbonic acid, or a reactive derivative thereof, to form the product esterified at 5-position.

After completion of the reaction sequence, the product formed is isolated from the reaction mixture in a conventional manner and, if desired, subjected to further purification in a conventional manner, for example by various chromatographic techniques, in particular column chromatography. In some cases, the compounds of formula (I) may be obtained in the form of product mixtures, but it is not always necessary to separate the individual compounds.

The starting compounds of formula (II) are known compounds or may be prepared by methods known in the art. As related literature, reference is made to U.S. Pat. British Patent Publication No. 57-120589 (Kokai), J. Am. Chem. Soc., 103, 4216 (1981), U.S. Patent 4,199,569. and 4,289,760. U.S. Patent Nos. 2,166,436; British Patent No. 215,654; European Patent Publication No. 904,709; Belgian Patent No. 32,034; Reissue U.S. Patent No. 4,457,920. U.S. Patent No. 2,615, and European Patent No. 2,615.

Conventional methods for converting compounds containing R ^{3 into} hydrogen may be used to convert R ^{3 to} compounds having any of the above groups. Preferably, the conversion is carried out prior to performing steps A and B. For example, methylation of the 5-hydroxy group is described in 237.341. can be carried out in the same manner as described in European Patent Specification No. 6,198.

The protection of the 5-hydroxy group with a protecting group such as a silyl group can be accomplished by reaction with a silyl halide in a solvent and preferably in the presence of a base.

There is no restriction on the solvent used for silylation, provided that it behaves inert during the reaction. Preferred solvents for this purpose include hydrocarbons such as hexane or benzene; chlorinated hydrocarbons such as methylene chloride or chloroform; ethers such as diethyl ether; and polar solvents such as tetrahydrofuran, acetonitrile, dimethylformamide, dimethylacetamide, dimethylsulfoxide or pyridine.

Examples of preferred parenting agents include Si (R ') (R ") (R"') Z, wherein R ', R "and R' 'are independently alkyl, benzyl or phenyl containing from 1 to 4 carbon atoms. and Z is a halogen ion or other suitable cation. Examples of preferred silylating agents include trialkylsilyl chlorides, triarylsilyl chlorides and silyl trifluoromethanesulfonates. Parenting is typically carried out in the presence of a base. The nature of the base is not essential, but in general organic bases are preferred over inorganic bases. Examples of useful bases include lithium sulfide, imidazole, pyridine, triethylamine and dimethylaminopyridine, especially imidazole.

Some silylating agents, such as hexamethyldisilazane and diethylrimethylsilamine, act simultaneously as a silylating agent and form their own base, so that they can be used with good results.

-6HU 198723 Β

The esterification of the 5-hydroxy group with a carboxylic acid or carbonic acid can be carried out as described in Step B above.

Methods of converting one group into another group are also known within the meaning of the group -X-Y-, as disclosed in European Patent 74758.

Naturally occurring starting materials 10 may be single compounds or mixtures of two or more compounds which can be used without isolation. Because, for example, mixtures of milbemycin A ₃ and A _{4 are} readily available and easy to use, such mixtures can be used to carry out the process of the invention without prior isolation.

Some of the compounds of formula (I) may also be prepared by fermentation.

Thus, the present invention further relates to a fermentation process for the preparation of compounds of formula I wherein -XY- is -CH ₂ -CH ₂ -, R ³ and R ^{4 are} hydrogen, and (i) R ¹ is is methyl and R ² is 2-methyl-1-propenyl; (ii) ^one R 25 is methyl and R ² is 2-methyl-lbutenilcsoport or (iii) ^R1 is ethyl and ^R2 is 2-methyl-1-propenyl.

Such compounds miibe- 30 tobramycin i »n (see opportunity to (i) above), will be I ₁₃ (see (ii) option above), and _{a] 4} (see an opportunity to (iii) above) the compounds hereinafter referred to as to call. These compounds are prepared by culturing a productive microorganism belonging to Streptomyces hygroscopicus species. For example, such a microorganism is the strain SANK 60286 belonging to the species Streptomyces hygroscopicus, which has been isolated from soil in the city of Miura (Kanagawa Prefecture, Japan) and its mutants and variants. 40

Milbemycin aj, oj ₃ and ₁₄ form part of the compounds isolated from the fermentation broth obtained by fermentation of strain SANK 60286, which are referred to as milbemycin ajj compounds and are identified in the following table.

R, R ₂

11 Methyl 2-methyl-1-propenyl 12 methyl isobutyl 13 methyl 2-methyl-l-enyl 14 ethyl 2-methyl-l-propenyl 15 ethyl isobutyl

Characteristics of Streptomyces SANK 60286

The mycological properties of the actinomycetes strain SANK 60286 are as follows.

1. Morphological characteristics

The strain undergoes microscopic branching of basal mycelia between pale yellow and yellowish brown, with white and yellowish-gray aerial micelles protruding with helical ends. In a spore colony or chain of spores, 10 or more spores can be observed in a line, and the spores have a wispy-grooved surface. In some types of media, the strain forms a clear golden mucus on the surface of the aerial mycelium, which turns into yellowish spots during fermentation. In later stages of cultivation, sometimes black spots are formed due to moisture.

2. Growth on different media

After culturing on different media for 14 days at 28 ° C, the strain exhibits the following characteristics. The color names and code numbers are based on the Guide to Color Standard, published by Nippon Shikisai Kenkyusho, Japan.

Media Feature ¹ ^

specific

Sucrose-nitrate G agar AM R SP Glucose-asparagine G agar AM R SP glycerol-asparagine G agar (ISP 5) AM R SP inorganic starch (i salt agar (ISP 4) AM R SP thyrotinic agar (ISP 7) G AM R SP

good, greyish-white (N-9) good, white-yellowish-gray not very good, greyish-gray (2-9-11) good, greyish-white, (N-9) pale yellow (3-9-10) no, yellowish-brown (2-9- 11) good, white-yellowish gray (2-9-11) pale yellow (8-9-11) not very good, pale olive (6-8-11) abundant, white-pale olive-gray (4-8-11) ) tan (2-9-11) no good, light olive gray (2-8-11) good, white to pale yellow (3-9-10) pale yellow (6-8-10) no

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Medium Properties Characteristic

peptone élesztőextraktum- G -agar agar (ISP 6) AM R SP nutriens agar {difco G company product AM R SP yeast extract, malt G extract agar (ISP 2) AM R SP oatmeal agar G (ISP 3) AM R SP water agar G AM SP burgonyakívonat-beet G extract agar AM SP

good, yellowish gray (2-9-12) good, grayish yellowish (4-9-11) no good, yellowish gray (1-9-10) poor formation, gray (N-9) pale yellow (3-9-10) nines very good, tan (2-9-11) abundant, greyish-white (N-9) red-yellow (12-8-9) pale yellow (8-9-12) very good, pale yellow (8-9-12) abundant, greyish-white ( N-9) tan-brown (2-9-11) pale olive-gray (4-7-11) faint, greyish-white (N-9) faint, pale brown-gray (2-8-8) no faint, greyish-white (N-9) ) good, light brownish gray (2-9-9) none

1) G: growth; AM: aerial mycelium; R: inverted surface; SP: soluble pigment

3) Physiological properties Physiological properties of SANK 60286 strain

the next ones: hydrolysis of starch positive liquefaction of gelatin positive reduction of nitrate (weak) positive milk coagulation (at 28 'C and 37' C) positive peptonization of milk (at 28'C and 37'C) (weak) positive growth temperature range (weak) 18-37 'C

(Medium ¹ ) ¹ ) Production of melanoid pigment

(Medium 2) negative (Medium 3) negative (Medium 4) negative 2) Medium 1: Yeast extract malt extract agar (ISP 2) Media 2: tripton - yeast extract medium (ISP 1) Medium 3: peptone -élesztőkivonat Iron Agar (ISP 6) Media 4: Tyrosine Agar (ISP 7)

At 14 ° C for 14 days on Pridhem-Gottlieb agar medium, SANK 60286 uses carbon sources as shown in the table below.

-8GB 198723 Β sugar recovery

D-glucose ++ L-arabinose ++ D-xylose + i-inositol ++ D-mannitol 4- + D-fructose ++ L-rhamnose ++ saccharose ++ raffinose ++ control -

++ good recovery; + recovery; - no recovery.

4. Cellular components

The cell wall of SANK 60286 was tested according to the method described by Becker, B., et al., Applied Microbiology, 12, 421 (1964). Since L, L-diaminopimellic acid and glycine could be detected, the cell wall can be considered as type 1 cell wall. Subsequently, the sugar components of whole cells were examined by Lechevalier, Μ. P., 1968, 71, 934 (Journal of Laboratory and Clinical Medicine). No characteristic distribution was found.

In summary, the strain was found to belong to the genus Slreplomyces belonging to the Actinomycetes.

Identification of the SANK 60286 strain is provided by the ISP (The International Streptomyces Project); Bergey's Manual of Determinative Bacteriology,

8th edition; Waksman, S. A. The Actinomycetes c. and other recent publications on Actinomycetes.

Comparison of the properties of SANK 60286 with those of other known strains of the Streptomyces genus shows that the morphological and physiological properties are nearly identical to those of Streptomyces hygroscopicus subsp. J. Antibiotics, 36, 438 (1983). aureolacrimosus.

However, a slight difference was observed between the breeding characteristics of the two strains. It is well-known that the characteristics of Aeliners change to some extent after some successive breeding. therefore, strains may not always be distinguished taxonomically due to minor differences in breeding characteristics. For these reasons, milbemycin produces substances αμ, «π» "13 ·" 14 "15

SANK 60286 Streptomyces hygroscopicus subsp. aureolaerimosus strain SANK 60286. In accordance with the provisions of the Budapest Agreement, the strain was deposited with the Fermentation Research Institute, Japan, on October 20, 1986, under the FERM BP-1190 accession number. Samples of the strain will be available in accordance with the relevant provisions of the Budapest Agreement.

Actinomycetes including Streptomyces hygroscopicus subsp aureolacrimus SANK

60286 strains are prone to mutation, both for natural reasons and for artificial treatments such as ultraviolet radiation, X-rays or chemical treatments. All productive mutants of strain SANK 60286 are included within the scope of the present invention. These mutant strains include mutants obtained by gene manipulation such as recombination, transduction and transformation. In other words, all strains capable of producing one or more of milbemycin, J3 and [4], and in particular strains which are not clearly distinguishable from SANK 60286 or its strains, are within the scope of the invention. mutánsaitól.

Milbemycin <μ, π and _[ alpha _{] 4 can be} prepared by culturing SANK 60286 in a suitable medium and isolating the milbemycins from the medium. Materials commonly used in the cultivation of microorganisms of the genus Streptomyces may be used as nutrients, such as glucose, sucrose, starch, glycerol, dense maltose syrup, molasses or soybean oil. Examples of sources of nitrogen include soybean meal, wheat germ, meat extract, peptone, yeast cells, corn jam, ammonium sulfate, and sodium nitrate. If desired, inorganic and organic additives such as calcium carbonate, sodium chloride, potassium chloride and phosphates can be used in appropriate combinations to promote the growth of microorganisms and activate the formation of milbemycin, π cs γ ₄ macrolides.

Similar to conventional methods of preparing antibiotics, the microorganism of the invention is cultured on a liquid medium, most preferably by submerged culture, under aerobic conditions. The culturing temperature is preferably from 22 ° C to 30 ° C, in most cases at about 28 ° C. In the case of shaking or tank breeding, milbemycin «11,« 13 and «| ₄ production typically reaches its peak after 5 to 15 days.

Production of milbemycin «μ,« ₁₃ and «ι _{4 is} as follows. A 1 mL sample of the culture was transferred to a small tube and 9 mL of 80% aqueous methanol was added. The tube is shaken and centrifuged. Subsequently, high performance liquid chromatography was performed using, for example, a 6 mm x 150 mm reverse phase column supplied by Senshu Co., Japan under the trade name H-2151 ODS, and a model sold by Hitachi, Japan under the Model 655. Five µl of the sample were injected into the column and run at a flow rate of 1.5 ml / minute with 80:20 acetonitrile / water. The appearance of milbemycin «μ,« 13 and u _{14 is} detected by an ultraviolet detector operating at 240 nm and is quantified, for example, by Union Tech. Inst. as defined by the MCPD - 350PC data processing unit.

-9HU 198723 Β

Various adsorbents, such as activated carbon, alumina, or silica gel, are collected from the culture to collect milbemycin ?,? And? ₄ ; synthetic adsorbents such as Diaion HP-20 (Japanese Mitsubishi Chem. Ind. Ltd.) adsorbents such as Avicel (Asahi Chem. Ind. Co., Ltd.) or filter paper: ion-exchange gel. based filter materials. What we do most effectively is:

The culture is filtered using a filter aid such as diatomaceous earth. The filter cake obtained is extracted with methanol to dissolve the desired compounds in aqueous methanol. To the resulting extract was added water and the aqueous mixture was extracted with hexane. The hexane solution is evaporated under reduced pressure to give an oily substance containing milbemycin ai, a3 and ai ₄ . This oily substance was then adsorbed on a Lobar Si60 column (size B, Merck, USA) and eluted with 8: 2 hexane: ethyl acetate to give fractions containing one product of milbemycin. , aj ₃ or ₁₄ . The fractions containing any of the milbemycins were then concentrated under reduced pressure to give an oily substance. A small amount of methanol was added to the oily substance and the resulting mixture was applied to a Lobar RP-8 column (size B, manufactured by Merck, USA). Elution is carried out with a 80:20 by volume mixture of acetonitrile and water. The fractions containing the individual compounds were then separated and the acetonitrile was evaporated under reduced pressure and extracted with ethyl acetate. Finally, high pressure liquid chromatography using a reversed phase column gives powdered milbemycin A 1, Q 1 3 and ₁₄ as separate powders. Milbemycin qjj and aj ₃ can be prepared in a similar manner.

The compounds of the present invention exhibit strong acaricidal activity against adult individuals and eggs of, for example, Tetranychus, Panonychus (e.g. Panonychus ulmi and Panonychus citri), and pests, which primarily kill fruit trees, vegetables and flowering plants. The compounds of the present invention are also effective against Ixodidae, Dermanyssidae and Sarcoptidae, which are parasites of animal species. The compounds of the present invention are also active in the so-called. exoparasites such as Oestrus, Lucilia, Hypoderma, Gautrophilus, lice and fleas, which breed parasitic life on animals and birds, especially domestic animals and poultry. The compounds of the present invention are also effective against domestic insects such as cockroaches and domestic air, as well as harmful insectivores parasitizing in agricultural and horticultural areas such as aphids and larvae of the Lepidoptera species. They are also effective against various species of soil, such as Meloidogyne, Bursaphelencus and Rhizoqlyphus.

The compounds of the invention also have excellent anti-parasitic activity against parasitic endoparasites in humans and animals. In particular, the compounds of the present invention are not only active against parasitic nematodes in domestic animals such as pigs, sheep, goats, cattle, horses, dogs, cats or poultry, but also in filamentous organs of the genus Filariidae and Setariidae. or effective against parasites in the blood.

If a composition comprising a compound of the present invention is to be used for agricultural or horticultural purposes, a variety of forms and compositions are possible. For example, the composition may be prepared in the form of dusts, coarse dusts, soluble powders, microgranules, fine microgranules, wettable powders, dilutable emulsions, emulsifiable concentrates, aqueous or oily suspensions, dispersions or solutions (which may be sprayed directly or diluted), aerosols or for example, the latter made with polymeric materials. In view of the nature of the carrier employed, it may be natural or synthetic and may be organic or inorganic, generally used to facilitate delivery of the active ingredient to the substrate to be treated and to facilitate storage, transport or handling of the active ingredient. Solid liquid gaseous carriers may be used, among those commonly used in compositions of this type.

Solid carriers which may be used include inorganic materials such as clays (e.g. kaolinite, montmorrilonite and attapulgil), talc, mica, pyrophyll, pumice, vermiculite, gypsum, calcium carbonate, dolomite, diatomaceous earth, magnesium carbonate, apatite, zeolite, and synthetic calcium silicate; organic materials of plant origin such as soybean meal, tobacco powder, wheat flour, walnut powder, wood flour, starch and crystalline cellulose; synthetic or natural high molecular weight materials such as coumarone resins, kerosene resins, alkyd resins, polyvinyl chloride, polyalkylene glycols, ketone resins, ester resins, copal resins and dammar resins; waxes such as carnauba wax and beeswax; and urea.

Suitable liquid carriers include paraffinic or naphthenic hydrocarbons such as kerosene, petroleum, spindle oil and white oil; aromatic hydrocarbons such as benzene, toluene, xylene, ethylbenzene, cumene and methylnaphthalene; chlorinated hydrocarbons such as carbon tetrachloride, chloroform, trichlorethylene, monochlorobenzene and 2-chlorotoluene; ethers such as dioxane and tetrahydrofuran; ketones, ex10

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EN 198723 such as acetone, methyl ethyl ketone, diisobutyl ketone, cyclohexanone, acetophenone and isophorone; esters such as ethyl acetate, amyl acetate, ethylene glycol acetate, diethylene glycol acetate, dibutyl maleate and diethyl succinate; alcohols such as methanol, n-hexanol, ethylene glycol, diethylene glycol, cyclohexanol and benzyl alcohol; ether alcohols, such as ethylene glycol phenyl ether, dielylene glycol ethyl ether and diethylene glycol butyl ether; polar solvents such as dimethylformamide and dimethylsulfoxide; and water.

Suitable gaseous carriers include air, nitrogen, carbon dioxide and freon. These gases can also be used mixed with spray formulations.

Various surfactants and high molecular weight compounds may be added, if desired, to enhance certain properties of the compositions, such as dispersibility, emulsifiability, distribution, permeation and adhesion, resulting in improved adhesion and absorption of the composition by the animal and plant. .

Surfactants for emulsifying, dispersing, wetting, spreading, binding, controlling degradation, stabilizing the active ingredients, enhancing fluidity and preventing rust formation may be of the nonionic, anionic, cationic, and amphoteric types, generally nonionic and / or anionic surfactants. Suitable nonionic surfactants include, for example, the polymerization adducts of long chain alcohols such as lauryl alcohol, stearyl alcohol or oleyl alcohol with ethylene oxide; polymerization adducts of alkylphenols such as isooctylphenol or nonylphenol with ethylene oxide; polymerization adducts of alkyl naphthols such as butyl naphthol or octyl naphthol with ethylene oxide; ethylene oxide polymerization adducts of long-chain fatty acids such as palmitic acid, stearic acid or oleic acid; polymerisation adducts of mono- or dialkylphosphoric acids such as stearylphosphonic acid or dilaurylphosphoric acid with ethylene oxide; polymerisation adducts of amines such as dodecylamine or stearinamine with ethylene oxide; esters of polyalcohols, such as long chain fatty acids of sorbitan, and polymerization adducts of these esters with ethylene oxide; and polymerization adducts of propylene oxide with ethylene oxide. Suitable anionic surfactants include, for example, alkyl sulfate (ester) salts, such as the amine salts of the sulfuric acid ester of sodium lauryl sulfate or of the oleyl alcohol; alkylsulfonals, such as the sodium salt of di-ethyl ester of succinic acid or the sodium salt of 2-ethylhexenesulfonic acid; and arylsulfonates, such as the sodium salt of isopropylnaphthalenesulfonic acid, the sodium salt of methylene bisnaphthalenesulfonic acid, the sodium salt of lignin sulfonic acid, and the sodium salt of dodecylbenzene sulfonic acid.

In order to improve the properties of said formulations and enhance their biological activity, the formulations in combination with high molecular weight compounds or other excipients such as casein, gelatin, albumin, soda, sodium alginate, carboxymethyl cellulose, methyl cellulose, hydroxyethyl cellulose or may be used in combination with polyvinyl alcohol.

The carriers and various excipients described above can be used alone or in a variety of combinations, for example, depending on the type of formulation and the intended use, among many other factors.

Dusts generally contain, for example, from 1% to 25% by weight of the active ingredient and the solid carrier and dispersant and wetting agent in the amount necessary for 100% by weight. If necessary, a protective colloid, thixotropic agent and an antifoam agent may be added.

Emulsifiable concentrates generally contain, for example, 5 to 50% by weight of active ingredient, 5 to 20% by weight of emulsifier and 100% by weight of a liquid carrier. If necessary, corrosion inhibitor may be added.

Oily formulations generally contain, for example, from 0.5 to 5% by weight of active ingredient and 100% by weight of a liquid carrier such as kerosene.

The aerosol will generally contain, for example, 0.1 to 5% by weight of active ingredient, optionally an odorant, and an amount of 100% by weight of an oily and / or liquid carrier. Liquid hydrocarbons, fluorocarbon gases and carbon dioxide can be used as the propellant gas.

Once prepared as described above, the compositions of the present invention can be usefully utilized in crop plants and domestic animals that are parasitic to harmful insects or mites. Plants can be treated in flooded areas, in the highlands and in orchards by treating the stems and leaves of the crop and the land around them. In both plant and animal treatments, the concentration of the active ingredient can be in the range of 0.5-100 ppm.

If the compounds of the invention are to be used as antelmintics in animals and humans, they may be administered orally in the form of a drink. The beverage is usually a solution or dispersion in a solution prepared with a suitable non-toxic solvent or water and a suitable suspending agent such as bentonite and a wetting agent or other excipient. Usually the beverage also contains an antifoam agent. The beverage will generally contain from 0.01 to 0.5% by weight, preferably from 0.01 to 0.1% by weight, of the active ingredient.

When the compounds of the present invention are to be administered to animals in the form of feed, the compounds are dispersed homogeneously in the feed, either as top dressing or in the form of pellets. The kfvan

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In order to provide an anthelmintic effect, the amount of active substance in the feed shall be between 0.0001 and 0.02% by weight.

The compounds of the invention may also be administered parenterally to animals in the anterior stomach, muscle or trachea, or by subcutaneous injection, dissolved or dispersed in a liquid vehicle. For parenteral administration, the active compounds are preferably admixed with a suitable vegetable oil, such as peanut oil or cottonseed oil. Formulations of this type generally contain from 0.05% to 50% by weight of the active ingredient.

The compounds of the invention may also be applied topically after mixing with a suitable carrier such as dimethylsulfoxide or a hydrocarbon type solvent. This type of formulation can be applied directly to the body surface of the animal by spray or direct injection.

The oral dose required to obtain the most favorable results will depend on a number of factors, such as the type of animal to be treated and the type and degree of parasitic infection. Generally, the dosage will be 0.01 to 100 mg, preferably 0.5 to 50 mg per kg body weight. The dose may be administered once or more times over a relatively short period of time, for example 1-5 days.

The following examples are intended to illustrate the invention.

Example 1

Milbemicin au, ₁₃ and βι ₄ (compounds 1, 2 and 3)

In ten 500-ml Erlenmeyer flasks, 100-100 ml of the pre-culture medium containing 1% sucrose, 0.35% polypeptone and 0.05% dipotassium hydrogen phosphate were inoculated with one inoculum of Streptomyces hygroscopicus subsp. aureolaerimosus strain SANK 60286 and cultured in a rotary shaker at 28 ° C for 48 hours. The 1 liter of culture fluid thus obtained is weighed into two 30-liter fermenters. Each contains 20 to 20 liters of medium containing 8% sucrose, 1% soybean powder, 1% skimmed milk, 0.1% yeast extract, 0.1% meat extract, 0.3% potassium carbonate, 0.03% dipotassium hydrogen -phosphate, 0.1% magnesium sulfate heptahydrate and 0.005% ferric sulfate heptahydrate and a pH of 7.2 before sterilization.

Cultivation was carried out at 28 ° C for 12 days in fermentors with a sterilized air flow of 0.5 volumes / well (wm) at an internal pressure of 0.5 kg / cm 2, 40-180 rpm and 4-7 ppm. At DO. After culturing, 32 liters of the fermentation broth is mixed with 1.8 kg of Cellte filtration aid and then filtered. The mycelial filter cake was washed with 5 liters of water, and the filtrate and washings were discarded. The mycelial filter cake was stirred with 20 liters of methanol for 1 hour and then washed again with 5 liters of methanol.

The filtrate and the washings were combined and concentrated under reduced pressure to give an aqueous residue of about 2 liters. This aqueous liquid was extracted with hexane (2 x 2 L) and the combined extracts were dried over anhydrous sodium sulfate, filtered and evaporated to give 38 g of an oily substance. Dissolve 20 g of this oily substance in 500 ml of hexane and apply the resulting solution to a column of 300 g of silica gel sold by Mallinckrodt, Inc. under the tradename Siliea Type 60 in hexane. Elution was carried out with 2 liters of hexane followed by 2 liters of a 3: 1 by volume mixture of hexane and ethyl acetate.

The eluted fractions were analyzed by high performance liquid chromatography (HPLC) to give αm, <* ^ ₂ , ^Q 13> ** 14 and ^{, J!} The substances are isolated as a mixture. Evaporation of the solvent gave 1.9 g of an oily crude product. The whole is dissolved in 10 ml of 50% aqueous methanol and the resulting solution is applied to a column which is treated with 160 g of 50% aqueous methanol from silica gel, marketed by Merck Co. under the tradename Art 7719. prepared. Elution was 60%, then 70%, then 80%. aqueous plugs containing methanol. The eluted fractions were analyzed by high performance liquid chromatography (SLC) to separate the mixture of milbemycin un, αι ₂ , 13, ₁₄ and 045.

Evaporation of the eluent gave 840 mg of an oily crude product. The whole is dissolved in 20 ml of acetonitrile and the resulting solution is subjected to preparative high pressure liquid chromatography on a reversed phase column, Senshu Co., Japan, ODS H-5251, 20 mm by 250 mm. In each case, 1 ml of the solution is applied to the column. Elution was carried out with an aqueous mixture of 80% acetonitrile at a flow rate of 9.9 ml / min. The fractions were analyzed by ultraviolet light at 240 mm to separate the fractions containing each product. The eluent from each fraction is then distilled off and the remaining aqueous residue is lyophilized. Thus, in each order of elution, 128 mg of milbemycin / l, 11.7 mg of milbemycin / ₂ , 14.8 mg of milbemycin / _{3 are in} powder form. ₍₄₃ mg of milbemycin ^a 14'l and 3 mg of milbemycin α 15 are obtained.

milbemicin α η

Analysis% (%): C, 68.83; H, 8.32.

Molecular weight: 626 (based on electron bombardment mass spectrum: this method will continue to be used).

C _{3 ()} H ₅ O ₀ >

Specific rotation: ²³ + + 104.3 ′ (c = 1.05, chloroform) (relative to the D-line of sodium herein and hereinafter).

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HU 193723 Β

UV Absorption Spectrum: _mex (CH ₃ OH) nm (E ^1% , cm -1): 230 (shoulder), 238 (990), 252 (shoulder).

IR absorption spectrum: ν _max (KBr) cm ^-1 : 3450, 2950, 1715, 1650, 1450, 1380, 1330, 1270, 1220, 1180, 1160, 1140, 1090, 1080, 1050, 1020, 990,940,850.

Proton Nuclear Magnetic Resonance Spectrum (CDCl ₃ , 270 MHz): 1.90 (3H, singlet, trans -CH ₃ -C (CH ₃ ) = CH-COO), 2.15 (3H, singlet, cis -CH ₃ -C (CH ₃ ) = CH-COO), 3.27 (1H, doublet of quartets, C 25 H), 4.00 (1H, doublet, J = 6 Hz, C ₆ H), 4.65-4.90 (4H, multiplet). , C ₂₆ H, C ₂₇ H).

Electrospray Mass Spec: m / z = 626 (M ⁺ ), 558, 526, 508, 400, 181, 153.

HPLC: Retention time: 13.4 minutes (Senshu Co. ODS H-2151 6.150 mm column: 80% acetonitrile / 20% water: 1.5 mL / min ultraviolet light detection) At 240 nm. These conditions continue to be used), milbemycin m?

Analysis% (%): C = 67.04,

H, 8.01.

M.W. 628Ősszegképlet: 36 ^ ^{Η 52θ9}

Specific rotation [u] ²³ = +118.3 '(c = 1.0, chloroform)

UV absorption spectrum: _max (CH ₃ OH) nm (E ^1%, cm '): 238 (750) 244 (810), 253 (shoulder)

IR absorption spectrum ν _max (KBr) cm ^-1 : 3500, 2950, 1740, 1710, 1450, 1380, 1330, 1290, 1180, 1120, 1090, 1050, 990.

Electrospray Mass Spec: m / z = 628, 556, 548, 525, 400, 382, 329, 181, 153.

HPLC retention time: 14.6 min on milbemycin

Analysis% (%): C = 67.41,

H, 8.12.

Molecular Weight: 640

Formula: C57H52O9

Specific rotation [a] ²³ = +91.6 ° (c = 0.89, CHC1 ₃₎

UV absorption spectrum: _max (CH ₃ OH) nm (E ^1%, cm '): 230 (sh), 237 (805) 245 (795), 253 (shoulder).

IR absorption spectrum: j / _max (KBr) ^cm-1: 3500, 2950, 1720, 1650, 1450, 1380, 1340, 1310, 1270, 1210, 1180, 1140, 1115, 1095, 1050, 990, 960.940, 860th

Proton Nuclear Magnetic Resonance Spectrum (CDCl ₃ , 270 MHz): 1.07 (3H, triplet, CH ₃ CH ₂ -C (CH ₃ ) = CH-COO), 2.16 (3H, singlet, CH ₃ -C (C ₂ H ₅ ) = CH-COO -, 3.27 (1H, doublet of quartets, C 1 H), 4.00 (1H, doublet, J = 6 Hz, C ₆ H), 4.65 - 4.90 ( 4H, multiplet, C ₂₆ H, C ₂₇ H).

Electrospray Mass Spec: m / z = 640 (M ⁺ ), 526, 508, 276, 181, 153.

HPLC retention time: 16.4 min. milbemicin α π

Analysis% (%): C, 67.62; H, 7.84

Molecular Weight: 650: C ₃₇ H _{J 2} O 9

Specific rotation [?] ²³ * = +96.1 '(c = 1.14, CHCl ₃ )

UV absorption spectrum: _max (CH ₃ OH) nm (E ^1%, cm ^'): 230 (sh), 237 (800), 244 (810), 253 (shoulder).

Proton Nuclear Magnetic Resonance Spectrum (CDCl ₃ , 270 MHz): 3.09 (1H, doublet of triplets, J = 2.4, 9.3Hz, C 25 H), 4.00 (1H, doublet, J = 6.6Hz, C ₆ H), 4.64-4.88 (4H, multiplet, C ₂ H, C ₂₇ H), 5.81 (1H, broad singlet, C ₃ H).

IR absorption spectrum: ν _max (KBr) cm ^-1 : 3450, 2950, 1720, 1650, 1450, 1380, 1340, 1270, 1220, 1180, 1140, 1100, 1060, 1030, 990, 960, 860.

Electrospray Mass Spec: m / Z = 640, 540, 522, 276, 263, 195, 167.

HLPC retention time: 18.0 min. milbemicin ακ Molecular Weight: 642 Molecular Formula: C ^ H ^ C ^

Electrospray Mass Spec: m / z =

642, 555, 540, 414, 396, 385, 256, 314, 264, 245, 195,167.

HPLC retention time: 19.6 min.

Example 2

5- (O-tert-Butyldimethylsilyl) -milbemycin a ₃₄ (Compound 13)

To a solution of 319.3 mg of 26-hydroxy-5- (O-tert-butyldimethylsilyl) -milbemycin / ¾ in 10 ml of methylene chloride cooled to 0 ° C was added 150 μ 150 of pyridine and 200 μΐ of 3-methyl- 2-butenoyl chloride and the resulting mixture was stirred for 30 minutes. The reaction mixture was poured into a saturated aqueous sodium bicarbonate solution and extracted with methylene chloride. The extract was dried over anhydrous magnesium sulfate and evaporated.

The residue was purified by silica gel column chromatography to obtain the title compound (326.2 mg, 91%).

Mass Spectrum (? Method, m / z): 754 (M ⁺ ), 736, 715, 697.654, 636, 597, 589.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl 3, δ ppm): 0.11 (3H, singlet, SiCH ₃ ), 0.13 (3H, singlet, SiCH ₃ ), 3.07 (1H, doublet of triplets, J = 2 4, 9.3Hz, C25 H), 3.85 (IH, d, J = 5.6Hz, C H _O), 5.78 (lH, singlet, C H _3).

Example 3

Milbemicin <»14 (Compound 3)

302.8 mg of 5- (O-tert-butyldimethylsilyl) -milbetnicin d | _To a cooled 0 ° C side of acetonitrile (15 mL) was added 2.5 mL of a 68% solution of hydrogen fluoride in pyridine, and the resulting mixture was stirred for 2.5 hours followed by potassium carbonate (about 300 mg). and adding water in the specified order. After extraction with chloroform, the extract was dried over anhydrous magnesium sulfate and evaporated. The residue was purified by silica gel column chromatography,

232.2 mg (90%) of the title compound having the properties of Example 113

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HU 198723 Β da produced milbemycin with the same properties as ₁₄ .

Example 4

26- (2-Butenoyl-o) - milbemycin Aj (Compound 5)

To a solution of 26-hydroxy-5- (O-tert-butyldimethylsilyl) -milbemycin A (150 mg) in methylene chloride (2 mL) cooled to 0 ° C was added pyridine (24 μ és) and 2-butenoyl chloride (39 μΐ). the resulting reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was poured into ice water and extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and evaporated. The residue was dissolved in 4 ml of acetonitrile and the resulting solution was cooled to 0 ° C. To the solution was then added 0.5 ml of a 68% solution of hydrogen fluoride in pyridine, and the resulting mixture was stirred at room temperature for 1.5 hours. The reaction mixture was poured into ice water and extracted with ethyl acetate. The extract was washed with a saturated aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate and evaporated. The residue was purified by preparative thin layer chromatography to give 66.5 mg (48%) of the title compound.

Mass spectrum (EI method, m / z): 626 (M ⁺ ), 540, 522, 414, 264, 245, 191, 167, 151.

Proton Nuclear Magnetic Resonance Spectrum 8270 MHz, CDCl ₃ , δ ppm): 1.99 (3H, doublet of doublets, J = 6.9, 1.6 Hz, CH-CH ₃ = CH-COO-), 3.08 (1H, doublet of triplets, J = 2,4,9,3Hz, C25 H), 4.84 (lH, d, J = 13.3Hz, C ₂₆ H), 5.87 (lH, quartet of doublets, J = 1.6, 15,3Hz, C H ₃ CH = CH-COO), 7.03 (lH, quartet of doublets, J - 6,9,15,3Hz, CH ₃ CH = CH-COO-).

Example 5

26- (3-Methyl-2-butenoyloxy) -milbemycin D (Compound 16)

To a solution of 177.3 mg of 26-hydroxy-5- (O-tert-butyldimethylsilyl) -milbemycin O and 0.1 ml of pyridine in 15 ml of methylene chloride is added 0.15 ml of 3-methyl-2 -butenoyl chloride and the resulting mixture was stirred for 2 hours. The reaction mixture was then poured into saturated aqueous sodium bicarbonate and extracted with methylene chloride. The extract was dried over anhydrous magnesium sulfate and evaporated. The residue was dissolved in acetonitrile (10 mL) and the resulting solution was cooled to 0 ° C and 1 mL of a 68% solution of hydrogen fluoride in pyridine was added. The reaction mixture was stirred for 2 hours and then poured into a saturated aqueous solution of sodium bicarbonate. After extraction with ethyl acetate, the extract was dried over anhydrous magnesium sulfate and evaporated. The residue was purified by silica gel column chromatography to obtain 144.7 mg (86%) of the title compound.

Mass spectrum (EI method, m / z): 654 (M ⁺ ), 618,554, 428,410,356.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 3.07 (1H, broad, doublet, J = 7.7Hz, C 1 H), 4.48 (1H, broad doublet, J 1 = 4, 0Ηζ, C ₅ H), 4.68 (1H, doublet, J = 14.7Hz, C ₂₆ H), 4.79 (1H, doublet, J = 14.7Hz, c _{2 (} , h).

Compounds listed below a

2-5. can be prepared according to the methods described in Examples.

Example 6

26- (2-Methyl-2-butenoyloxy) -milbemycin A (Compound 6)

Mass spectrum (EI method, m / z): 640 (M +), 604.540, 522.414.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 1.80 (3H, doublet, J = 7.2Hz, CH ₃ -CH = C (CH ₃ ) -COO-), 1.85 (3H, singlet, CH ₃ -CH = C (CH ₃ ) -COO-), 3.08 (1H, doublet of triplets, J = 2.4, 9.3Hz, C 1 H), 4.73 (1H, doublet, J-13 , 7Hz, C ₂₆ H).

Example 7

26- (2-Hexenoyloxy) -milbemycin A ₄ (Compound 7)

Mass spectrum (EI method, m / z): 654 (M ⁺ ), 618, 522, 504, 414, 396.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 0.93 (3H, triplet, J = 7.3 Hz, CH-CH ₃ CH ₂ CH = CH ₂ -COO-), 4.02 (1H , doublet, J = 13.3Hz, C ₂₆ H), 4.86 (1H, doublet, J = 13.3Hz, C ₂₆ H), 7.01 (1H, triplet of doublets, J = 6.9.15, 7Hz, C ₃ H _r CH = CH-COO-)

Example 8

26- (4-Methyl-2-pentenoyloxy) -milbemycin A (Compound 8)

Mass Spectrum (EI method, m / z): 654 (M +), 618, 522.414, 396.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 1.07 (6H, doublet, J = 6.9Hz, (CH ₃ ) ₂ CH-CH = CH-COO), 3.08 (1H, triplets) doublets, J = 2.4, 9.3 Hz, C? H), 4.72 (lH, d, J = 13.3Hz, C ₂₆ H), 4.86 (lH, d, J = 13.3Hz, C ₂₆ H), 6.99 (1H, triplet of doublets, J = 6.5, 15.7Hz, (CH ₃ ) ₂ CH-CH-CH-COO -).

Example 9

26- (3-Hexanoyloxy) -milbemycin Aj (Compound 9)

Mass spectrum (EI method, m / z): 654 (M +), 414, 279, 195, 167.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 3.07 (2H, doublet, J-6.4Hz, CH-C), 4.68 (1H, doublet, J-12.9Hz, C26H) , 4.79 (1H, doublet, J = 12.9Hz, C26H)

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Example 10

26- (4-Pentenoyloxy) -milbemycin A4 (Compound 10)

Mass spectrum (Method E1, m / z): 640 (M ⁺ ), 604, 522, 264, 195.167.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl ₃ , δ ppm): 3.07 (1H, doublet of triplets, J = 2.4, 9.3Hz, C25H), 4.69 (1H, doublet, J = 13.7Hz) , C26H), 4.79 (1H, doublet, J = 13.7Hz, C26H), 4.99-5.10 (2H, multiplet, CH2 = CH-CH2CH2-COO-).

Example 11

26- (Cinnamoyloxy) -milbemycin A4 (Compound 11)

Mass Spectrum (EI method, m / z): 688 (M +), 652, 522, 276, 195, 167.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCB, δ ppm): 3.07 (1H, doublet of triplets, J = 2.4, 9.7Hz, C25H), 4.81 (1H, doublet, J = 13.3Hz) C26H), 4.89 (1H, doublet, J = 13.3Hz, C26H), 6.47 (1H, doublet,

J = 16.1Hz, Ph-CH = CH-COO), 7.72 (1H, doublet, J = 16.1Hz, Ph-CH = CH-COO-).

Example 12

26- (4-Chloro-cinnamoyloxy) -milbemycin A4 (Compound 12)

Mass Spectrum (E1 method, m / z): 722 (M +), 704, 540, 522.504.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl3, ppm): 3.07 (1H, doublet of triplets, J = 2.4, 8.9Hz, C25H), 4.81 (1H, doublet, J = 13.3Hz, C26H) ), 4.93 (1H, doublet, J = 13.3Hz, C26H), 6.44 (1H, doublet,

J = 16.1Hz, 4-Cl-Ph-CH-CH-COO-, 7.67 (1H, doublet, J = 16.1Hz, 4-Cl-Ph-CH = CH-COO).

Example 13

5- (O-Propionyl) -2- (3-methyl-2-butenoyloxy) milbemycin A4 (Compound 14)

Mass Spectrum (? Method, m / z): 696 (M ⁺ ), 604, 504, 414, 396, 356, 264, 195,167,151.

Proton Nuclear Magnetic Resonance Spectrum: (270 MHz, CDCl 3, δ ppm): 1.15 (3H, triplet, j = 7.6Hz, CH ₃ CH ₂ CO-), 2.40 (2H, quartet, J = 7.6Hz) , CH 3 CH 2 CO-), 3.07 (1H, broad triplet, J = 8.0Hz, C ₂₅ H), 4.11 (1H, doublet, J = 6.0Hz, C 6 H) k, 4.50 - 4.76 (4H, multiplet, C ₂₆ H, C ₂₇ H), 5.6. -5.95 (5H, multiplet, C ₃ H, C ₅ H, C ₉ H, C ₁₀ H, (CH ₃ ) ₂ C-CH-COO).

Example 14

5- (O-Ethoxycarbonyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4 (Compound 15)

Mass Spectrum (EI method, m / z): 712, (M ⁺ ), 414, 396, 264, 195, 167, 151.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl3, δ ppm): 1.31 (3H, triplet, J = 7.1Hz, CH3CH2OCOO-), 3.07 (1H, doublet of triplets, J = 2.4, 8.9Hz) , C 25 H), 4.14 (1H, doublet, J = 6.1 Hz, C ₆ H), 4.19 (2H, quartet,

J = 7.1Hz, CH3CH2OCOO-), 4.57 to 4.76 (4H, m, ₂ C EH, C ₂₇ H), 5.54 (lH, doublet of doublets, J = 1,6,6,1Hz, 5H).

Example 15

26- (3-Methyl-2-butenoyloxy) -ivermectin B _ta (Compound 17)

Mass spectrum (FAB method, triethanolamine added, m / z): 1014, 992, 978, 962, 934, 878,830, 299.194.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl 3, δ ppm): 3.07-3.29 (3H, multiplet, C ₄ H, C / H, C ₂₅ H), 3.35 (1H, singlet, C ₂ H), 3.48 (6H, singlet, 2-OCH ₃ ), 3.97 (1H, doublet, J = 6.1 Hz, C ₆ H), 4.49 (1H, broad singlet, C ₅ H), 4.69 ( 2H, broad singlet, C ₂₇ H),

4.97 (1H, broad doublet, J = 7.3 Hz, C ₁₅ H).

Example 16

26- (3-Methyl-2-butenoyloxy) S-541A (Compound 20)

Mass spectrum (Method E1, m / z): 710 (M ⁺ ), 592, 523, 468, 448, 423, 375.

Proton Nuclear Magnetic Resonance Spectrum (270 MHz, CDCl 3, δ ppm): 3.75 (1H, doublet, J = 10.9Hz, C ₂₅ H), 4.67 (1H, doublet, J = 13.3Hz, C ₂₆ H), 4.82 (1H, doublet, J = 13.3Hz, C ₂₆ H), 5.20 (1H, doublet, J = 8.9Hz, C ₃₂ H).

Example 17

Activation test on adult mites

0.3, 1 or 3 ppm. Concentrate test solutions of the compounds of the present invention and one of the following three reference compounds: Mixture of C 1 and C ₂ of milbemycin, prepared as described in U.S. Pat. Japanese issue number! known from the document; 26-acetoxy-milbemycin A4; and 26-acetoxy-avermectin B] _a . The solutions also contain 0.01% of a spreading agent.

The cotyledons of cow pea plants (Vigna sinensis Savi) are inoculated with an organic phosphorus-type insecticide-active two-spotted spider mite (Tetranychus urticae). One day after inoculation, 7 ml of one of the above-mentioned experimental solutions are sprayed with a rotary sprayer manufactured by Mizuho Seisakusho Co., Japan, to provide a utilization rate of 3.5 mg / cm ² per leaf surface. After spraying, the leaves are allowed to stand at 25 ° C. After 3 days, the binocular microscope is used to determine whether or not the adult insects have died and the percentage mortality is calculated. The results are given in the table below.

Number of compound 3 ppm mortality 1 ppm 0.3 ppm ¹ («ll) 100 100 93.0 2M 100 95.6 77.3 3 (a 14) 100 100 100 5 100 100 54 6 100 100 55 7 100 100 61

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HU 198723 Β

8 100 100 51 9 100 100 50 10 100 100 44 11 100 100 40 12 100 100 55 14 100 100 65 15 100 100 60 16 100 100 57 17 100 100 53 20 100 100 59

Example 18

Activity assay on mite eggs

Experimental solutions of 1 or 3 ppm were prepared using the same reference compounds as described in Example 17.

Female adult spider mites are allowed to ovulate on cow pea cotyledons and the adult mites are removed to obtain experimental leaves containing about 50 to 50 eggs.

The experimental solution is then applied to the leaves as described in Example 17. After storage at 25 ° C for two weeks, the number of non-hatching eggs is determined and the percentage of non-hatching eggs is calculated.

The results are given in the following table.

The compound has an ovicidal activity (%) of 3 ppm to 1 ppm

¹ («ll) 94 51 2 (an) 94 71 3 (ai ₄ ) 95 57 5 94 45 6 62 33 7 77 50 8 55 23 10 53 23 14 82 41 15 63 20 16 67 21 19 43 20 Ci + C ₂ 2.7 2.1 26-acetoxy- 31 12 -milbemicin A4 26-Acetoxy- 4.2 2.4 B _la -averraektin

From the tables above it can be seen that the new milbemycins according to the invention have very strong acaricidal activity against adult mites at concentrations as low as 0.3 ppm, but they are also valuable ovicidal.

The pesticides of the present invention may be in the form commonly used for this purpose, may contain conventional carriers and other excipients, or may be prepared by conventional means, for example in the following compositions.

parts

Wettable powder formulation

Example 3 Compound 95 Sodium Lignin Sulfonate 3 Sodium Methylene Bisnaphthalene Sulfonate 2

Emulsifiable Concentrate Example 1 Compound 40 xylene polyoxyethylene nonylphenyl ether 10 calcium dodecylbenzenesulfonate 5

Oily preparation

Example 20 Compound 5 5 kerosene 80 methyl naphthalene 15

dusts

Example 16 Compound 3 Clay 60 Talc 36.5 Isopropyl hydrogen phosphate 0.5

Claims (14)

PATENT CLAIMS A process for the preparation of a novel macrolide compound of formula (I): wherein -XY- represents -XY- represents -CH ₂ -CH ₂ - or CH ₂ -CH ₂ H-, R ¹ is methyl, ethyl, isopropyl, sec-butyl or -C (CH ₃ ) = CHR ⁵ and in the latter R ³ is methyl, ethyl or isopropyl, R ² is - (CH 2) n C) R ⁶ ) = C (R ⁷ ) (R ⁸ ) wherein n is 0, 1 or 2, R ⁶ and R ⁷ independently of one another are hydrogen or methyl; R ⁸ is hydrogen or C 1-4 alkyl, or phenyl optionally substituted with halo, provided that when R ⁸ is C 1-4 alkyl, then - (CH 2) _n C (R ⁶ ) = C (R ⁷ ) the alkenyl group of the formula R ^{8 has} up to 8 carbon atoms, R ³ is hydrogen, C 2 -C 5 alkanoyl, C 2 -C 5 alkoxycarbonyl, or tri (C 1 -C 4) alkylsilyl, and R ⁴ is hydrogen or alpha-L-oleandrosyl-alpha-L-oleandrosyloxy, with the proviso that R ⁴ is hydrogen when R ¹ is methyl, ethyl or -C (CH ₃ ) = CHR ⁵ ; in the latter, ^R5 is methyl, ethyl or isopropyl - preparation, characterized in that a) a compound of formula (III): wherein -XY-, R ¹ , R ³ and R 4 are as defined herein, with an acid of formula R ² COOH, wherein R ² is as defined herein, or any of or a reactive derivative thereof; or b) milbemycin A 1, milbemycin a; _{To produce 3} or ₁₄ , the producing microorganism belonging to the species Strcptomyees hygroscopicus is cultured and the desired product is isolated. -161 HU 198723 Β Process (a) according to claim 1, characterized in that the starting material is a compound of the formula (III) in which -XY- is -CH ₂ -CH ₂ -, -CH ₂ CHOH- or -CH ₂ -. C (= O) -, while R ¹ , R ³ and R ⁴ are as defined in claim 1. The method according to a) or Claim 2 3. The first claim, characterized in that the starting material used is (111) the compound wherein ^R¹ is methyl, ethyl or isopropyl and R ⁴ is hydrogen . Process according to claim 1 (a) or claim 2, characterized in that the starting material is a compound of formula (III) in which R ¹ is isopropyl or sec-butyl and R ⁴ is alpha-L-. oleandrosyl-alpha-L-oleandrosyl group. The process of claim 1 a) or claim 2; characterized in that starting material is a compound of formula (III) wherein R ¹ is -C (CCH ₃ ) = CHR ⁵ , wherein R ⁵ is as defined in claim 1 and R ⁴ is hydrogen. 6. A method according to claim 1 or 2 to 5. A process according to any one of claims 1 to 6 for the preparation of compounds of formula I wherein R ² is -CH = C (CH ₃ ) (R ¹⁰ ) wherein R ¹⁰ is methyl or ethyl. R ² is a carboxylic acid of formula COOH or a reactive derivative thereof. 7. The method according to claim 1, a) or 2-6. A process for the preparation of compounds of formula I according to any one of claims 1 to ³ , wherein R ³ is hydrogen, alkanoyl containing 2 to 5 carbon atoms or alkoxycarbonyl having 2 to 5 carbon atoms, characterized in that the corresponding compound of formula III starting material. A process according to claim 1 for the preparation of compounds of formula I wherein -XY- is -CH ₂ -CH ₂ , R 'is methyl or ethyl, R ² is 2-melyl-1- propenyl, while R ³ and R ⁴ are hydrogen, wherein appropriate starting materials are used. A process according to claim 1 for the preparation of the following compounds of formula (I): 26- (3-Methyl-2-butenoyloxy) -milbemycin A ₃ 26- (3-Methyl-2-pentenoyloxy) -milbemycin A ₃ 26- (3-Methyl-2-butenoyloxy) -milbemycin Aj 26- (3-Methyl-2-pentenoyloxy) -milbemycin A _{4 3} 26- (2-Butenoyloxy) -milbemycin A4 26- (2-Methyl-2-butenoyloxy) -mylbemycin A4 26- (2-Hexenoyloxy) -milbemycin A4 26- (4-Methyl-2-pentyloxy) -mylbemycin A4 26- (3-Hexenoyloxy) -milbemycin A4 26- (4-Pentenoyloxy) -milbemycin A4 26- (Cinnamoyloxy) -milbemycin A4 26- (4-Chloro-cinnamoyloxy) -milbemycin A ₄ 5- (O-tert-Butyl-dimethylsilyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4 5- (O-Propionyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A4 5- (O-Ethoxycarbonyl) -2- (3-methyl-2-butenoyloxy) milbemycin A ₄ 26- (3-Methyl-2-butenoyloxy) -milbemycin D 26- (3-Methyl-2-butenoyloxy) -ivermectin B _1a 26- (3-Methyl-2-butenoyloxy) avermectin B _1a 23-Deoxy-26- (3-oxy-methyl-2-butenoyloxy) S-541A 26- (3-methyl-pyran-2-bulenoil) -S-541A 23-Oxo-26- (3-methyl-2-butenoyloxy) -S-541A, characterized in that the appropriate starting compounds are used. A process according to claim 1 for the preparation of the following compounds of formula (I): 26- (3-Methyl-2-butenoyl-yloxy) milbemycin A ₃ 26- (3-Methyl-2-pentenoyloxy) -milbemycin A ₃ 26- (3-Methyl-2-butenoyloxy) -milbemycin A4 26- (3-Methyl-2-pentenoyloxy) -milbemycin A4 26- (2-Hexenoyloxy) -milbemycin A4 26- (4-Methyl-2-pentenoyloxy) -milbemycin A4 26- (Cinnamoyloxy) -milbemycin A4 5 - (() - propionyl) -2- (3-methyloxy-2-buienoil) - milbemycin z \ ₄ 5- (O-Ethoxycarbonyl) -2- (3-methyl-2-butenoyloxy) -milbemycin A ₄ 26- (3-Methyl-2-butenoyloxy) -milbemycin D 26- (3-Methyl-2-butenoyloxy) -ivermectin B _1a 26- (3-Melyl-2-butenoyloxy) avermectin Bj _a 23-Deoxy-26- (3-oxy-methyl-2-butenoyloxy) S-541A 26- (3-Methyl-2-butenoyloxy) S-541A 23-Oxo-26- (3-methyl-2-butenoyloxy) -S-541A, characterized in that the appropriate starting compounds are used. A process according to claim 1 for the preparation of the following compounds of formula (I): 26- (3-Methyl-2-butenoyloxy) -milbemycin A ₃ 26- (3-Methyl-2-pentenoyloxy) -milbemycin A ₃ 26- (3-Methyl-2-butenoyloxy) -mibemycin A4 26- (3-Methyl-2-pentenoyloxy) -milbemycin A4 26- (3-Methyl-2-butenoyloxy) -milbemycin D 26- (3-Methyl-2-butenoyloxy) -ivermectin Bj _a 26- (3-Methyl-2-butenoyloxy) avermectin Bj _a 23-Deoxy-26- (3-oxy-methyl-2-butenoyloxy) S-541A 26- (3-Methyl-2-butenoyloxy) S-541A 23-Oxo-26- (3-methyl-2-butenoyloxy) -S-541A, characterized in that the appropriate starting compounds are used. Method b) according to claim 1, characterized in that the producing microorganism is Streptomyces hygroscopicus subsp. aureolacrimosus strain SANK 60286 (accession no .: FERM BP-1190). 13. A process for the preparation of an anti-parasitic pharmaceutical composition, wherein a macrolide compound of formula (I), wherein the substituents are as defined in claim 1, is a carrier and a pharmaceutically acceptable carrier for human or veterinary use. optionally mixed with other additives and made into an anti-parasitic composition. -171 HU 198723 Β 14. An anti-parasitic pesticide comprising 0.3 ppm to 95% by weight of a macrolide compound of formula I: wherein: -XY- represents -XY- represents -CH ₂ -CH ₂ - or CH 1 -CHOH-, R ¹ is methyl, ethyl, isopropyl, sec-butyl or -C (CH ₃ ) = CHR ⁵ and in the latter is methyl, ethyl or isopropyl. R ⁷ is - (CH 2) n C) R ⁶ ) = C (R ⁷ ) (R ⁸ ) wherein n is 0, 1 or 2, R ⁶ and R ⁷ independently of one another are hydrogen or methyl; R ⁸ is hydrogen or C 1-4 alkyl or phenyl optionally substituted with halo, provided that when R ⁸ is C 1-4 alkyl, then - (CH 2) _n -C (R ⁶ ) = C ( R ⁷ ) The alkenyl group of the formula (R ⁸ ) contains up to 8 carbon atoms, R ³ is hydrogen, C 2 -C 5 alkanoyl, C 2 -C 5 alkoxycarbonyl, or tri (C 1 -C 4 alkylsilyl), and R ⁴ is hydrogen or alpha-L-oleandrosyl-alpha-L-oleandrosyloxy, with the proviso that R ⁴ is hydrogen when R ¹ is methyl, ethyl or -C (CH ₃ ) = CHR ⁵ and in the latter R ⁵ is methyl, ethyl-propyl or propyl, and is present in an amount of 100% by weight of solid or liquid organic or inorganic origin , containing natural or artificial inert carriers, preferably clay, talc, xylene, kerosene and anionic and / or nonionic surfactants, preferably emulsifying, dispersing or suspending agents.